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Antibodies
Volume 8
Issue 4
10.3390/antib8040049
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Article

The Impact of Immunoglobulin G1 Fc Sialylation on Backbone Amide H/D Exchange

by
Felix Kuhne
1,2,
Lea Bonnington
1,
Sebastian Malik
1,
Marco Thomann
1,
Cecile Avenal
3,
Florian Cymer
3,
Harald Wegele
1,3,
Dietmar Reusch
1,
Michael Mormann
2 and
Patrick Bulau
1,*
1
Pharma Technical Development, Roche Diagnostics GmbH, Nonnenwald 2, 82377 Penzberg, Germany
2
Institute of Hygiene, University of Muenster, Robert-Koch-Strasse 41, 48149 Muenster, Germany
3
Pharma Technical Development Analytics Biologics, F. Hoffmann-La Roche Ltd., 4070 Basel, Switzerland
*
Author to whom correspondence should be addressed.
Antibodies 2019, 8(4), 49; https://doi.org/10.3390/antib8040049
Submission received: 30 August 2019 / Revised: 24 September 2019 / Accepted: 26 September 2019 / Published: 1 October 2019
(This article belongs to the Special Issue Higher Order Structure Characterization of Therapeutic Antibodies)
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Abstract

The usefulness of higher-order structural information provided by hydrogen/deuterium exchange-mass spectrometry (H/DX-MS) for the structural impact analyses of chemical and post-translational antibody modifications has been demonstrated in various studies. However, the structure–function assessment for protein drugs in biopharmaceutical research and development is often impeded by the relatively low-abundance (below 5%) of critical quality attributes or by overlapping effects of modifications, such as glycosylation, with chemical amino acid modifications; e.g., oxidation or deamidation. We present results demonstrating the applicability of the H/DX-MS technique to monitor conformational changes of specific Fc glycosylation variants produced by in vitro glyco-engineering technology. A trend towards less H/DX in Fc Cγ2 domain segments correlating with larger glycan structures could be confirmed. Furthermore, significant deuterium uptake differences and corresponding binding properties to Fc receptors (as monitored by SPR) between α-2,3- and α-2,6-sialylated Fc glycosylation variants were verified at sensitive levels.
Keywords: hydrogen/deuterium exchange; mass spectrometry; Fc glycosylation; antibody conformation; higher-order structure; biopharmaceutical; antibody effector function; FcγR binding; structure-function; sialic acid linkage hydrogen/deuterium exchange; mass spectrometry; Fc glycosylation; antibody conformation; higher-order structure; biopharmaceutical; antibody effector function; FcγR binding; structure-function; sialic acid linkage

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MDPI and ACS Style

Kuhne, F.; Bonnington, L.; Malik, S.; Thomann, M.; Avenal, C.; Cymer, F.; Wegele, H.; Reusch, D.; Mormann, M.; Bulau, P. The Impact of Immunoglobulin G1 Fc Sialylation on Backbone Amide H/D Exchange. Antibodies 2019, 8, 49. https://doi.org/10.3390/antib8040049

AMA Style

Kuhne F, Bonnington L, Malik S, Thomann M, Avenal C, Cymer F, Wegele H, Reusch D, Mormann M, Bulau P. The Impact of Immunoglobulin G1 Fc Sialylation on Backbone Amide H/D Exchange. Antibodies. 2019; 8(4):49. https://doi.org/10.3390/antib8040049

Chicago/Turabian Style

Kuhne, Felix, Lea Bonnington, Sebastian Malik, Marco Thomann, Cecile Avenal, Florian Cymer, Harald Wegele, Dietmar Reusch, Michael Mormann, and Patrick Bulau. 2019. "The Impact of Immunoglobulin G1 Fc Sialylation on Backbone Amide H/D Exchange" Antibodies 8, no. 4: 49. https://doi.org/10.3390/antib8040049

APA Style

Kuhne, F., Bonnington, L., Malik, S., Thomann, M., Avenal, C., Cymer, F., Wegele, H., Reusch, D., Mormann, M., & Bulau, P. (2019). The Impact of Immunoglobulin G1 Fc Sialylation on Backbone Amide H/D Exchange. Antibodies, 8(4), 49. https://doi.org/10.3390/antib8040049

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